Protein Complex for Intracellular Delivery via Segmentation
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Solution Overview
Problem
Current methods for delivering biologically active macromolecules, such as proteins and peptides, into cells are hindered by cell membrane permeability issues, stability concerns, and limited targeting ability, leading to low efficacy and instability in vivo.
Innovation Solution
A protein complex comprising an in vitro stabilization protein like ubiquitin, a membrane translocation sequence domain, a biologically active molecule, an in vivo stabilization protein such as serum albumin binding peptide, and a nucleus-cytoplasm signal domain, which facilitates efficient intracellular delivery and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If biologically active macromolecules (proteins, peptides) are used as therapeutic agents, then physiological selectivity and efficacy are improved, but cell membrane permeability deteriorates
Solution Approach 1:
The protein is divided into functional domains: a cell permeable domain (first region) that facilitates membrane crossing, and a biologically active domain (second region) that performs the therapeutic function inside the cell. This segmentation allows the protein to overcome the membrane barrier while maintaining its biological activity.
Solution Approach 2:
The first region acts as an intermediary or delivery vehicle that mediates the transport of the biologically active second region across the cell membrane. This intermediary domain enables the macromolecule to penetrate the membrane without compromising the activity of the functional region.
2Ease of operation
If delivery systems using short peptides are developed, then cell permeability is improved, but in vitro and in vivo stability deteriorates
Solution Approach 1:
The invention creates a composite protein structure combining a cell permeable domain with a biologically active domain. This composite design integrates the membrane-crossing capability of short peptides with the stability and functionality of full-length proteins, achieving both permeability and stability.
Solution Approach 2:
The invention merges the beneficial characteristics of short peptides (cell permeability) with those of stable proteins (structural integrity and activity). By combining these two functional elements into a single protein construct, the invention achieves both delivery capability and stability.
3Reliability
If biologically active macromolecules are delivered into cells, then therapeutic efficacy is improved, but delivery efficiency deteriorates due to cell membrane barrier
Solution Approach 1:
The protein is segmented into a delivery-specific first region and a functional second region. This segmentation ensures that the delivery function is optimized for membrane penetration while the functional region maintains high therapeutic efficacy, thereby improving overall delivery efficiency.
Solution Approach 2:
The invention modifies the structural parameters of the protein by introducing specific domains with optimized properties for membrane interaction. The first region has parameters (amino acid sequence, structure) specifically tuned for efficient membrane crossing, enhancing delivery efficiency.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The protein complex effectively delivers biologically active molecules into cells, enhancing their stability and functionality, as demonstrated by its ability to suppress cancer cell growth in various cell lines, while maintaining solubility and avoiding immunogenicity.
Implementation Method 1
a membrane translocation sequence domain, a biologically active molecule
Implementation Method 2
an in vitro stabilization protein that is a ubiquitin (Ub), a Ub-like protein
Implementation Method 3
an in vivo stabilization protein that is a serum albumin binding peptide
Data Source
Figure 1
Figure 2(a)~2(b)
Figure 3
AI summary
A protein complex for intracellular delivery and a use thereof. The protein complex for intracellular delivery and a method of delivering a biologically active molecule into a cell by using the protein complex enable an effective delivery of a biologically active molecule.